Journal: Journal of Cancer

Article Title: SKA2-mediated transcriptional downregulation of the key enzyme of CoQ 10 biosynthesis PDSS2 in lung cancer cells

doi: 10.7150/jca.79058

Figure Lengend Snippet: Functional implication of SKA2-PDSS2 axis in lung cancer. ( a ) Boxplots of PDSS2 expression in TCGA lung cancer dataset. ( b ) PDSS2 expression levels were analyzed with qRT-PCR using OriGene TissueScan lung cancer panels (HLRT103). The stages were divided into I, II, III, and IV. “N” means normal. The wide bars denote the mean levels of PDSS2, while the narrow ones represent standard deviation away from the mean. ( c ) Survival analysis in relation to SKA2 and PDSS2 expression in lung cancer. Kaplan-Meier plot of overall survival of lung cancer patients in the GSE13213 dataset, the p-values were calculated using Log-rank test. ( d ) PDSS2 abrogates SKA2-promoted lung cancer cells growth. Cells were transfected with indicated plasmids and cell growth was continuously monitored by JULI Stage Real-time Cell History Recorder. ( e, f ) Cell motility analysis. Cell motility was also captured every 15 minutes for 12 hours by the aforementioned instrument. Motile trajectory, mean total migration distances and mean migration speeds are presented.

Article Snippet: TissueScan lung cancer panels (HLRT103) containing prepared cDNAs were purchased from OriGene (Rockville, MD).

Techniques: Functional Assay, Expressing, Quantitative RT-PCR, Standard Deviation, Transfection, Migration

Journal: Journal of Clinical Investigation

Article Title: The T-box transcription factor Brachyury promotes epithelial-mesenchymal transition in human tumor cells

doi: 10.1172/jci38379

Figure Lengend Snippet: Figure 1 Brachyury alters EMT marker expression and induces a highly migratory and invasive phenotype in human epithelial tumor cells. (A) Bright- field images of PANC-1-pcDNA and PANC-1–pBrachyury cells grown on a plastic surface (top 2 panels: original magnification, ×10) and immunofluorescence analysis of EMT markers in cells grown on cover glasses (bottom panels: original magnification, ×40) are shown. The green signal represents the staining of the corresponding protein, and the blue signal represents the DAPI-stained nuclei. (B) Western blot analysis of Brachyury and EMT markers. β-Actin is shown as the protein loading control. (C) Real-time PCR analysis of cDNA from the above cells for Brachyury and EMT markers. (D) In vitro cell migration and ECM invasion assays. Results of 1 of 3 experiments are shown. Error bars indicate SEM of triplicate measurements. *P < 0.05, **P < 0.001, ***P < 0.0001 for pcDNA versus pBrachyury.

Article Snippet: Real-time PCR was performed for Brachyury and GAPDH as described above on commercially available normal and tumor lung tissue cDNA panels (TissueScan Lung Cancer Tissue qPCR Arrays I and III, OriGene Technologies Inc.).

Techniques: Marker, Expressing, Immunofluorescence, Staining, Western Blot, Control, Real-time Polymerase Chain Reaction, In Vitro, Migration

Journal: Journal of Clinical Investigation

Article Title: The T-box transcription factor Brachyury promotes epithelial-mesenchymal transition in human tumor cells

doi: 10.1172/jci38379

Figure Lengend Snippet: Figure 6 Brachyury inhibition reduces tumor metastasis. (A) H460 cells stably transfected with con.shRNA (circles) or Br.shRNA vectors (triangles) were assessed for cell proliferation and survival by MTT assay. Three independent experiments showed comparable results; results from 1 experiment are shown. ***P = 0.0003 for con.shRNA versus Br.shRNA. Western blot shows cyclin D1 and β-actin expression. (B) Subcutaneous tumors were induced by injection of 4 × 106 of H460 cells in HBSS admixed with 50% (v/v) Matrigel. Graph shows volumes from 8 tumors at day 15 after tumor implantation. (C) cDNA from tumors in B were analyzed by real-time PCR for indicated markers. (D) Lungs from animals bearing subcutaneous tumors were collected at day 15 after tumor implantation, homogenized, and cultured in puromycin-containing medium. Graph shows visible colony counts. (E) Mice were inoculated with 7.5 × 105 H460 cells transfected as indicated via tail vein. Forty-five days after tumor implantation, animals were euthanized, and lungs were evaluated for tumor nodules. Graph shows results from 3 independent experiments. Experiments 1–3 are denoted by black, gray, and white circles (con.shRNA) and triangles (Br.shRNA), respectively. Statistical difference between treatment groups was analyzed by pooled results of the above experiments. Two representative lungs from each group are shown for comparison. White outlines and black arrowheads indicate tumor masses.

Article Snippet: Real-time PCR was performed for Brachyury and GAPDH as described above on commercially available normal and tumor lung tissue cDNA panels (TissueScan Lung Cancer Tissue qPCR Arrays I and III, OriGene Technologies Inc.).

Techniques: Inhibition, Stable Transfection, Transfection, shRNA, MTT Assay, Western Blot, Expressing, Injection, Tumor Implantation, Real-time Polymerase Chain Reaction, Cell Culture, Comparison

Journal: Journal of Clinical Investigation

Article Title: The T-box transcription factor Brachyury promotes epithelial-mesenchymal transition in human tumor cells

doi: 10.1172/jci38379

Figure Lengend Snippet: Figure 7 Brachyury expression in human lung tumor tis- sues. Real-time PCR was performed for Brachyury on lung tumor tissue cDNA from 80 lung cancer patients of the indicated stages of disease. The stage II, III, and IV cDNA samples are further rep- resented by squares, triangles, and ×’s, respec- tively. As controls, 16 samples of normal lung cDNA were also analyzed, each obtained from a pathologically normal section of lung from a can- cer patient. All values and the medians for each group are expressed as a ratio to the endogenous control GAPDH.

Article Snippet: Real-time PCR was performed for Brachyury and GAPDH as described above on commercially available normal and tumor lung tissue cDNA panels (TissueScan Lung Cancer Tissue qPCR Arrays I and III, OriGene Technologies Inc.).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Control

Journal: Cell death and differentiation

Article Title: PON3 is upregulated in cancer tissues and protects against mitochondrial superoxide-mediated cell death.

doi: 10.1038/cdd.2012.35

Figure Lengend Snippet: Figure 2 PON3 is found overexpressed in several human tumors. (a) PON3 cDNA expression levels were determined in cancer versus normal tissues by qRT-PCR using TissueScan cancer survey panels (covering B380 cDNA samples from various tumors) and (b) lung cancer panels (covering 24 matched samples, that is, diseased and healthy tissue from the same patient). The 24 samples were either summarized (‘total’) or separated into different groups of age, stage or diagnosis. (c) PON3 protein is found upregulated in human ovarian cancer tissue. Western blot analysis of PON3 in protein extracts from three independent normal ovary tissue (‘N’) and ovarian carcinoma (‘C’). GAPDH served as loading control

Article Snippet: PON3 expression levels were determined in cancer versus normal tissues by quantitative real-time PCR using TissueScan Cancer Survey Panel-III (4 96-well format) normalized to b-actin and GAPDH as described before.15 In addition, we determined PON3 cDNA levels in 24 samples from patient-matched TissueScan matched Lung Cancer Panel-IV (Origene, Rockville, MD, USA).

Techniques: Expressing, Quantitative RT-PCR, Biomarker Discovery, Western Blot, Control